Type 2 diabetes (T2D) mellitus is a chronic metabolic disease that afflicts over 500 million people worldwide and causes nearly 1.5 million deaths each year. Diabetics are hospitalized at a more frequent rate and have more expensive hospital visits compared to the general population. Current antibiotic-based treatments for infection contribute to the global rise in antimicrobial resistance and cause dysbiosis of the endogenous microbiome, leading to an adverse susceptibility to pathogens. Thus, additional strategies to prevent and cure bacterial infection are desperately needed. The vaginal microbiome is a key reservoir for pathogen dissemination to the urinary tract and a guardian against translocation to distant body sites by maintaining epithelial integrity and controlling inflammation. Group B Streptococcus (GBS) is a bacterium that is frequently detected in the vaginal microbiome without symptoms, but it is also an under-reported cause of aerobic vaginitis. In the diabetic population, which is 95% type 2 diabetics, GBS causes invasive disease in ~6,000 diabetic women annually and disproportionately causes urinary tract infections. However, neither the T2D vaginal microbiome nor its influence on diabetic susceptibility to this pathogen have been studied. Previous research suggest that the gut microbiome can respond to metabolic disease; in some cases diabetes is associated with enrichment of opportunistic pathogens in the gut or dysbiotic gut bacteria that are vulnerable to pathogens. In the context of pregnancy, gestational diabetes is associated with enriched abundance of opportunistic pathogens in the vaginal microbiome. This proposal aims to understand whether metabolic disease impacts the vaginal microbiome outside of pregnancy and whether microbiota mediate diabetic susceptibility to GBS. We hypothesize that T2D patients will have an altered microbiome and metabolite composition and an inflammatory cytokine profile, and that diabetic-derived communities will be less protective against GBS and concurrently inflict damage and inflammation on the host epithelium. This hypothesis will be tested by the following specific aims: 1) Define homeostatic characteristics of the vaginal microenvironment in T2D, and 2) Characterize T2D vaginal microbiome interactions with GBS and mediation of pathogen-host interactions. To complete these aims, innovative approaches such as miniature bioreactor cultivation and human vaginal stem-cell derived organoids will be used. The research will provide the applicant with training in organoid models, transcriptomics and metabolomics, and whole genome and 16S analysis, while building upon the applicant’s experience with the miniature bioreactor model. This training will take place at Baylor College of Medicine with mentorship from leading experts in metabolic disease, host-microbe interactions, human microbiome, and both proposed model systems, miniature bioreactors and organoids. This research will identify compon...